四环素胶体金免疫检测技术的研究  

Study on the Colloidal Gold Immunoassay of Tetracycline

作　　者：栗慧 金艳丹 张岩蔚 魏东 

机构地区：[1]河北省农产品安全检测重点实验室,河北张家口075000 [2]河北北方学院食品安全研究中心,河北张家口075000

出　　处：《食品研究与开发》2018年第3期146-150,共5页Food Research and Development

基　　金：河北省自然科学基金项目(C2016405007);河北省科技支撑计划项目(13227110D);河北省高校百名优秀创新人才支持计划(Ⅲ)(SLRC2017019);张家口市科技攻关计划项目(1611045C);河北北方学院重大项目(ZD201403);2017年河北省研究生创新资助项目(CXZZSS2017151)

摘　　要：为研制四环素(tetracycline,TC)胶体金免疫层析试纸条,对胶体金的制备及与四环素多克隆抗体的标记进行研究。采用柠檬酸钠还原法制备胶体金,向加热沸腾的100 mL0.01%氯金酸中加入1.8 mL1%柠檬酸钠,加热15 min后冷却定容;已制备好的胶体金溶液标记四环素抗体,将装有1 mL胶体金的离心管中加入一定量K_2CO_3调节p H值,加入一定量抗体蛋白后4℃条件下反应30 min,加入100μLNa Cl后4℃条件下保存备用,标记成功后优化条件。结果表明,胶体金颜色为紫红色、清亮透明、无沉淀及漂浮物,可见光光谱峰宽较小,证明胶体金大小分布均匀,制备成功;硝酸纤维素膜(NC膜)上有明显红色斑点,可见光光谱中最大吸收峰出现偏移,证明金标抗体标记成功;胶体金标记四环素抗体的最佳p H值为7.5,最佳标记蛋白量为稀释抗体6 000倍。胶体金的制备及与四环素多克隆抗体标记的成功,为装配四环素胶体金试纸条奠定了基础,为四环素在食品样品中快速检测方法的建立提供理论参考。In order to prepare colloidal gold immunochromatographic of tetracycline, the preparation of colloidal gold and colloidal gold labeled with tetracycline of polyclonal antibody were studied. Using sodium citrate reduction method to prepare colloidal gold, added 1.8 mL1 % C6H5Na3O7·H2O to boiling 100 mL0.01 % HAuCl4,cooling constant volume when after heating 15 min. The colloidal gold labeled with tetracycline of polyclonal antibody.A centrifuge tube with 1 mL colloidal gold added a certain amount of K2CO3 that regulate the pH value, after adding a certain amount of antibody protein, reaction of 30 min below 4 ℃, adding 100 μL NaCl preserve under 4 ℃.Optimization marked after the success. The results showed that the colloidal gold color was purple red, clear and transparent, no sediment and floating debris, the visible light spectrum width was small, the results showed that colloidal gold uniform, successfully prepared; nitrocellulose membrane （NC membrane） had obvious red spots, the largest visible spectrum the absorption peak shifted, that gold labeled antibody markers; colloidal gold labeled pH study built antibody was 7.5. The best labeled protein was 6 000 times the dilution antibody, colloidal gold and preparation of polyclonal antibody labeled with tetracycline, laid the foundation for the assembly of tetracycline colloidal gold test strip for rapid detection method tetracycline in food samples to provide theoretical reference.

关 键 词：四环素(TC)多克隆抗体 胶体金 金标抗体 

分 类 号：TS207.5[轻工技术与工程—食品科学]